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At least 451 records · Page 25Linked to original sources
PubMed2005

Elucidation of the monoclonal antibody 5G8-reactive, virulence-associated lipopolysaccharide epitope of Haemophilus influenzae and its role in bacterial resistance to complement-mediated killing.

The phase-variable locus lex2 is required for expression of a Haemophilus influenzae lipopolysaccharide (LPS) epitope of previously unknown structure. This epitope, which is reactive with monoclonal antibody (MAb) 5G8, has been associated with virulence of type b strains. When strain RM118 (from the same source as strain Rd), in which the lex2 locus and MAb 5G8 reactivity are absent, was transformed with lex2 DNA, transformants that were reactive with MAb 5G8 were obtained. Surprisingly, the 5G8 reactivity of these transformants was phase variable, although the lex2 locus lacked tetrameric repeats and was constitutively expressed. This phase variation was shown to be the result of phase-variable expression of phosphorylcholine (PCho) such that MAb 5G8 reacted only in the absence of PCho. Structural analysis showed that, compared to RM118, the lex2 transformant had acquired a tetrasaccharide, Gal-alpha1,4-Gal-beta1,4-Glc-beta1,4-Glc-beta1,4, linked to the proximal heptose (HepI). A terminal GalNAc was detected in a minority of glycoforms. LPS derived from a mutant of RM7004, a virulent type b strain which naturally expresses lex2 and has LPS containing the same tetrasaccharide linked to HepI as the sole oligosaccharide extension from the inner core, confirmed that GalNAc is not a part of the MAb 5G8-reactive epitope. Thus, MAb 5G8 specifically binds to the structure Gal-alpha1,4-Gal-beta1,4-Glc-beta1,4-Glc-beta attached via a 1,4 linkage to HepI of H. influenzae LPS, and we show that the ability to synthesize this novel tetrasaccharide was associated with enhanced bacterial resistance to complement-mediated killing.

Antibodies, Monoclonal↗
PubMed2006

Colorimetric method for phase evaluation.

Measurements of very small phase changes in optical measurement techniques are usually performed with interferometric methods that are based on evaluation of interference patterns, which correspond to a phase change of the investigated wave field. If values of the phase change are small, it is difficult to determine accurately the phase values, and one needs very expensive measurement systems. We present a simple method for evaluation of small phase variations that uses the interference of polychromatic light. The phase change affects the color of the interference pattern, and the color of the interference pattern corresponds to a specific phase change that can be evaluated using colorimetric analysis. We describe and analyze our colorimetric phase evaluation method. The proposed method offers accurate results, and it may be suitable for practical utilization in the optical industry.

Journal Article↗
PubMed2000

Lipid profiles of Helicobacter pylori colony variants.

A phase variation in Helicobacter pylori has been previously described. In one phase the bacterium had a cell wall lipid content typical for gram-negative bacteria (HpL), whereas in the other phase the bacterium was found to have a cell wall with increased amounts of lysophospholipids (HpS). The conversion is spontaneous, but could also be induced by acid (HpS(ind)) and was associated with in vitro release of Vac A and urease. The purpose of the present study was to determine the full phospholipid content of the cell wall to indicate a molecular mechanism of the colony variation. There were no appreciable differences between the lipid profiles of HpS and HpS(ind), while there were major differences between HpL and the S-variant. In the S-variant, lysophospholipids constituted about 50% of the total phospholipids, as compared to less than 2% in the L-variant. The proportion of total and individual cholesteryl glucosides also showed considerable changes. HpL was dominated by the phosphate-linked cholesteryl glucoside (72%) while the acylated cholesteryl glucoside was the main cholesteryl glucoside of the S-variant (65%). Our results demonstrate a dramatic change in cell wall properties after acid induction and spontaneously in vitro, and suggest some molecular mechanisms for this variation from an in vitro non-virulent to a virulent variant.

Cardiolipins↗
PubMed1996

Questions about gonococcal pilus phase- and antigenic variation.

Pathogenic organisms inhabit one of several defined locations within a host where temperature, pH, and nutrients are relatively constant. While the microorganism must adapt to different environments within the host, the host immune system is the most formidable predator that can limit the growth of a pathogen. Neisseria gonorrhoeae (the gonococcus, Gc) is the causative agent of gonorrhoea, and has evolved several systems for varying the antigenicity of different surface antigens, presumably to help evade the effects of the human immune system. The On/Off/On phase variation of surface structure expression also alters the antigenic characteristics of the bacterial cell surface. Antigenic variation of the major subunit of the pilus, pilin, occurs by unidirectional, homologous recombination between a silent locus and the expression locus. The silent loci lie from 1 to 900 kb from the expression locus in the chromosome yet all can donate their sequences to the expression locus. The genetic composition of the pilin loci of two Gc strains has been elucidated, and the types of changes that lead to altered forms of the pilus have been extensively characterized. However, little is known about the precise molecular mechanisms used to allow high-frequency, non-reciprocal, chromosomal recombination between pilin loci or about what regulates the process of maintaining chromosome fidelity.

Antigenic Variation↗
PubMed2006

Mycoplasma genitalium P140 and P110 cytadhesins are reciprocally stabilized and required for cell adhesion and terminal-organelle development.

Mycoplasma genitalium is a human pathogen that mediates cell adhesion by a complex structure known as the attachment organelle. This structure is composed of cytadhesins and cytadherence-associated proteins, but few data are available about the specific role of these proteins in M. genitalium cytadherence. We have deleted by homologous recombination the mg191 and mg192 genes from the MgPa operon encoding the P140 and P110 cytadhesins. Molecular characterization of these mutants has revealed a reciprocal posttranslational stabilization between the two proteins. Loss of either P140 or P110 yields a hemadsorption-negative phenotype and correlates with decreased or increased levels of cytoskeleton-related proteins MG386 and DnaK, respectively. Scanning electron microscopy analysis reveals the absolute requirement of P140 and P110 for the proper development of the attachment organelle. The phenotype described for these mutants resembles that of the spontaneous class I and class II cytadherence-negative mutants [G. R. Mernaugh, S. F. Dallo, S. C. Holt, and J. B. Baseman, Clin. Infect. Dis. 17(Suppl. 1):S69-S78, 1993], whose genetic basis remained undetermined until now. Complementation assays and sequencing analysis demonstrate that class I and class II mutants are the consequence of large deletions affecting the mg192 and mg191-mg192 genes, respectively. These deletions originated from single-recombination events involving sequences of the MgPa operon and the MgPa island located immediately downstream. We also demonstrate the translocation of MgPa sequences to a particular MgPa island by double-crossover events. Based on these observations, we propose that in addition to being a source of antigenic variation, MgPa islands could be also involved in a general phase variation mechanism switching on and off, in a reversible or irreversible way, the adhesion properties of M. genitalium.

Adhesins, Bacterial↗
PubMed1990

Coregulation of type 12 M protein and streptococcal C5a peptidase genes in group A streptococci: evidence for a virulence regulon controlled by the virR locus.

Group A streptococci express at least two surface-associated virulence factors, the antiphagocytic M protein and the antichemotactic streptococcal C5a peptidase (SCP). Preliminary evidence suggested that the biosynthesis of these two proteins is coordinately controlled and subject to simultaneous phase variation. To explore this possibility further, a series of phase-switching and phase-locked M- variants were assayed for SCP by enzyme-linked immunosorbent assay inhibition and for SCP-specific mRNA by dot blot hybridization. All M- cultures produced diminished amounts of SCP antigen and specific mRNA, whereas revertants produced quantities equivalent to those of the wild-type M+ culture. A phase-locked strain that harbors a deletion in a region upstream of the M12 and SCP genes, termed the virR locus, failed to produce SCP antigen or SCP-specific transcripts. The SCP-specific transcript produced by M+ bacteria was shown by Northern (RNA) blot hybridization to be 4 kilobases in size, distinguishing it from the transcript which encodes M protein. These data demonstrate that phase switching of both SCP and M12 proteins is at the transcriptional level and that expression is under the control of the upstream virR locus. We propose that the genetic determinants of these proteins and of colony morphology comprise a virulence regulon.

Adhesins, Bacterial↗
PubMed1998

Absence of seasonal variation in the phase of the endogenous circadian rhythm in humans.

Humans may be subject to seasonal variations, as evidenced by the existence of seasonal affective disorder (SAD) and midwinter insomnia. However, some recent studies have shown that the seasonal variation in the phase of the circadian rhythm is relatively weak in healthy humans. In the present study, evidence is found that there is no seasonal variation in the phase of the endogenous circadian rhythm at all. Body temperature, cortisol excretion, and subjective alertness of six subjects recorded under constant routine conditions showed no systematic seasonal variation in circadian phases. This finding indicates that secondary zeitgebers blocked or counterbalanced the seasonal variation in the entrainment effect of the natural photoperiod. The human being may live in an environment in which the photoperiod has lost its status of primary zeitgeber.

Adolescent↗
PubMed2004

Separation of B6 vitamers with micellar liquid chromatography using UV and electrochemical detection.

Separation of six vitamers of vitamin B6 was performed by RP-HPLC using micellar mobile phase, UV and electrochemical detection. Effect of temperature, type and amount of organic modifier in mobile phase on efficiency and asymmetry factor showed that, the appropriate conditions were temperature of 35 degrees C and 3.0-5.0% (v/v) 1-butanol in mobile phase. Variations of selectivity factor versus 1-butanol concentration, pH of mobile phase, and SDS concentration was investigated and the following optimized conditions were selected for the separation: 3.0% (v/v) 1-butanol, pH=5.5 and 65 mM SDS in mobile phase. Electrochemical behavior of vitamers in optimized mobile phase was investigated using cyclic voltammetry, and potential of +1.2 V versus Ag/AgCl(Sat.) was chose as working potential. Finally, separation of B6 vitamers using UV detection at 254 nm and electrochemical detection at +1.2 V was compared.

1-Butanol↗
PubMed1986

Overlapping deletion in two spontaneous phase variants of Coxiella burnetii.

Chromosomal DNA from the Nine Mile phase I strain of Coxiella burnetii (CB9MIC7) was cloned into the cosmid vector pHC79. The resulting gene library was probed with a radiolabelled HaeIII fragment present in the parental strain but absent from a spontaneously derived Nine Mile phase II strain (CB9MIIC4). The insert, which includes the missing HaeIII fragment, was 38.5 kb in length. When DNA from this cosmid clone was hybridized to genomic DNA of the parental CB9MIC7 and its derivative CB9MIIC4, a number of fragments were missing or altered in the latter strain. Restriction mapping localized the fragments to a contiguous portion of the chromosomal DNA fragment. The data were consistent with an 18 kb deletion in the chromosome of CB9MIIC4. Another intrastrain spontaneous derivative, CB9MI514, also lacked the sentinel HaeIII fragment and carried a deletion of approximately 29 kb within the same cloned insert. Both deletions appeared to share a common terminus, within the limits of resolution. In all other strains investigated, both phase I and phase II, the DNA represented by the insert seemed intact. The strains examined were representative of various stages of phase variation. The relationship between the observed deletions and the mechanism of phase transition in Nine Mile strains is discussed.

Chromosome Deletion↗
PubMed2002

The N-acetyltransferase RimJ responds to environmental stimuli to repress pap fimbrial transcription in Escherichia coli.

In uropathogenic Escherichia coli, P pili (Pap) facilitate binding to host epithelial cells and subsequent colonization. Whereas P pili can be produced at 37 degrees C, the expression of these fimbriae is suppressed at 23 degrees C. Previously, insertion mutations in rimJ, a gene encoding the N-terminal acetyltransferase of ribosomal protein S5, were shown to disrupt this thermoregulatory response, allowing papBA transcription at low temperature. In this study, we created an in-frame deletion of rimJ. This deletion relieved the repressive effects not only of low temperature but also of rich (Luria-Bertani [LB]) medium and glucose on papBA transcription, indicating that RimJ modulates papBA transcription in response to multiple environmental stimuli. papI transcription was also shown to be regulated by RimJ. papBA transcription is also controlled by a phase variation mechanism. We demonstrated that the regulators necessary to establish a phase ON state--PapI, PapB, Dam, Lrp, and cyclic AMP-CAP-are still required for papBA transcription in a rimJ mutant strain. rimJ mutations increase the rate at which bacteria transition into the phase ON state, indicating that RimJ inhibits the phase OFF-->ON transition. A DeltarimJ hns651 mutant is viable on LB medium but not on minimal medium. This synthetic lethality, along with transcriptional analyses, indicates that RimJ and H-NS work through separate pathways to control papBA transcription. Mutations in rimJ do not greatly influence the transcription of the fan, daa, or fim operon, suggesting that RimJ may be a pap-specific regulator. Overexpression of rimJ under conditions repressive for papBA transcription complements the DeltarimJ mutation but has little effect on transcription under activating conditions, indicating that the ability of RimJ to regulate transcription is environmentally controlled.

Acetyltransferases↗
PubMed1996

Application of the principle of optical phase-contrast microscopy to velocity phase-encoded MRI of blood flow in the aorta.

A new method of presenting magnetic resonance phase information is described and an example of its application given in the context of velocity phase-encoded MRI of blood flow in the aorta. The method takes as its starting point Zernike's technique of phase contrast microscopy. It exploits the parallel between the transform plane in Fourier optics and kappa-space in MRI. In the example described two datasets are acquired, one with and the other without velocity encoding, as in conventional phase-encoded velocity imaging. A dataset is formed which is corrected for unwanted phase variations caused by static field inhomogeneity. The method then effectively combines phase and magnitude information into a single image. The technique is complementary to existing methods of displaying phase information.

Aorta↗
PubMed1986

Novel regulatory mutants of the phosphate regulon in Escherichia coli K-12.

New pleiotropic mutants were isolated that express either the phoA, psiE or psiO promoter constitutively and simultaneously alter bacterial alkaline phosphatase regulation, carbon utilization or ultraviolet light sensitivity. To do this, Lac+ mutants were isolated from strains with the appropriate lacZ transcriptional fusions. Over 300 independent mutants were characterized, and all that constitutively express phoA map in phoR, phoU, the phosphate-specific transport system or a new locus called phoF. However, only phoU mutants express both phoA and psiE constitutively. Carbohydrate-utilizing mutants that show constitutive expression of psiE and psiO map in cya, crp and, possibly, crr. Also, numerous ultraviolet-light-sensitive mutants were discovered that show increased psiO expression and map in lon. Some other mutations that lead to constitutive psiO expression (which is normally induced either by phosphate, nitrogen or carbon starvation or anoxia) show decreased expression of phoA. Also, several mutants were found that show an unusual metastable character affecting psiO or phoA transcription. In these, colonies spontaneously switch between an induced and repressed "state" with respect to lac or bacterial alkaline phosphatase expression. In some, the clonal variation of the lactose phenotype or bacterial alkaline phosphatase synthesis is recA-independent and phenotypically resembles phase variation in Salmonella typhimurium. The latter class are called "phase mutants". The mutants are discussed in terms of protein-nucleic acid interactions and/or possible changes in the DNA, i.e. modifications or rearrangements, within the phosphate gene system, that are physiologically regulated.

Alkaline Phosphatase↗
PubMed2006

Static susceptibility effects in balanced SSFP sequences.

The steady state of balanced steady-state free precession (b-SSFP) sequences in presence of field inhomogeneities can be expressed in terms of a rapidly convergent series, which contains the characteristic function of the susceptibility related phase variations. As long as the intravoxel standard deviation sigma of the accumulated phase per repetition time (TR) is of order 1 (radians) or lower, the signal at TE=TR/2 behaves like a spin echo (SE), in accordance with recent observations. The signal decays quadratically around the maximum near TR/2 with a curvature, which solely depends on sigma and not on T'2. For larger sigma, a qualitative change toward a gradient echo (GRE) occurs and the signal shows characteristics of an usual FID. With increasing off-resonance the width of the SE regime decreases.

Algorithms↗
PubMed1998

The filamentous hemagglutinin of Bordetella parapertussis is the major adhesin in the phase-dependent interaction with NCI-H292 human lung epithelial cells.

Bordetella parapertussis is a Gram-negative bacterium which colonizes the human respiratory tract and can cause whooping cough or pertussis. This pathogen is subject to phase variation and expresses a series of virulence factors exclusively in the Bvg+ phase. Here, it is demonstrated for the first time that only the Bvg+ phase of B. parapertussis adheres to and invades the human lung epithelial cell line NCI-H292. A B. parapertussis mutant defective in expression of the Bvg+-regulated filamentous hemagglutinin (FHA) showed reduced binding (77% reduction) to NCI-H292 cells, as did a FHA mutant of the related Bordetella pertussis (85% reduction). In contrast to B. pertussis, binding of B. parapertussis to NCI-H292 cells was not inhibited by heparin, suggesting differences in the FHA adhesin and its host-cell receptor between these two species. Thorough understanding of the mechanism of action of the B. parapertussis virulence factors, such as FHA, is of particular interest in the development of novel strategies of pertussis vaccination.

Adhesins, Bacterial↗
PubMed2000

DNA methylation-dependent regulation of pef expression in Salmonella typhimurium.

Plasmid-encoded fimbriae (Pef) expressed by Salmonella typhimurium mediate adhesion to mouse intestinal epithelium. The pef operon shares features with the Escherichia coli pyelonephritis-associated pilus (pap) operon, which is under methylation-dependent transcriptional regulation. These features include conserved DNA GATC box sites in the upstream regulatory region as well as homologues of the PapI and PapB regulatory proteins. Unlike Pap fimbriae, which are expressed in a variety of laboratory media, Pef fimbriae were expressed only in acidic, rich broth under standing culture conditions. Analysis of S. typhimurium grown under these conditions indicated that Pef production was regulated by a phase variation mechanism, in which the bacterial population was skewed between fimbrial expression (phase ON) and non-expression (phase OFF) states. Leucine-responsive regulatory protein (Lrp) and DNA adenine methylase (Dam) were required for pef transcription. In contrast, the histone-like protein (H-NS) and the stationary-phase sigma factor (RpoS) repressed pef transcription. Methylation of the pef GATC II site appeared to be required for pef fimbrial expression based on analysis of a GCTC II mutant that did not express Pef fimbriae. Analysis of the DNA methylation states of pef GATC sites indicated that, under acidic growth conditions, which induced Pef production, most GATC I sites were non-methylated, whereas GATC II and GATC X were predominantly methylated. The methylation protection at GATC I and GATC II was dependent upon Lrp and was modulated by PefI. Together, these results indicate that Pef production is regulated by DNA methylation, which is the first example of methylation-dependent gene regulation outside of E. coli.

Base Sequence↗
PubMed1986

Bacterial alkaline phosphatase clonal variation in some Escherichia coli K-12 phoR mutant strains.

Several phoR alleles (phoR19, phoR20, phoR68, phoR69, phoR70, and phoR78) led to either a bacterial alkaline phosphatase (BAP)-constitutive phenotype or a variable behavior, depending upon the strain tested. Whereas Escherichia coli K10, MC1000, and XPh4 phoR mutants were constitutive, AB1157, BD792, MC4100, and W3110 phoR mutants displayed the metastable character. For the latter strains, constitutive mutants regularly segregated BAP-negative clones which yielded constitutive variants again at a high frequency. Indeed, the pattern of variation observed in BAP-variable phoR strains is phenotypically analogous to phase variation of the H1/H2 flagellum antigen type in Salmonella typhimurium and the molecular switch between the immune and sensitive states in bacteriophage lambda. The metastable behavior was not a general property of BAP-constitutive mutants, since several phosphate-specific transport-phoU mutations led to a constitutive (stable) phenotype regardless of the strain tested. But in phoR phosphate-specific transport-phoU mutants, the metastable character was epistatic (dominant), and such double mutants showed clonal variation in BAP-variable strains.

Alkaline Phosphatase↗
PubMed1964

INDUCTION BY ANTIBIOTICS AND COMPARATIVE SENSITIVITY OF L-PHASE VARIANTS OF STAPHYLOCOCCUS AUREUS.

Molander, C. W. (Cedars of Lebanon-Mount Sinai Hospitals, Los Angeles, Calif.), B. M. Kagan, H. J. Weinberger, E. M. Heimlich, and R. J. Busser. Induction by antibiotics and comparative sensitivity of L-phase variants of Staphylococcus aureus. J. Bacteriol. 88:591-594. 1964.-The penicillins, cephalothin, vancomycin, and bacitracin were found to be less inhibitory to the L-phase variants than to their respective parent bacteria. Those antibiotics not considered to be primarily inhibitors of cell-wall synthesis were, in general, somewhat more inhibitory to the L form than to their parent bacteria. Only the penicillins and cephalothin readily induced L-phase variation. Novobiocin induced pleomorphic growth resembling the earliest stages of L-phase transformation. Failure of observable induction by bacitracin and vancomycin suggests that these two antibiotics affect cell-wall synthesis in a manner different from the penicillins, or that L-phase transformation may require more than "penicillin-like" interference with cell-wall synthesis.

Anti-Bacterial Agents↗
PubMed1987

Improving MR image quality in the presence of motion by using rephasing gradients.

Numerous techniques exist for suppressing ghosting artifacts due to respiratory motion on MR images. Although such methods can remove coherent ghosting artifacts, motion during gradient pulses also leads to poor image quality. This is due to phase variations at the echo caused by changes in velocity from one phase-encoding view to the next. The effect becomes severe for long sampling times and long TE values and can lead to low estimates of T2. We discuss general, robust modifications of the standard gradient or spin-echo sequences by using rephasing gradients that force the phase of constant-velocity moving spins to be zero at the echo. These sequences lead to a significant reduction in motion artifacts and hence improvement in image quality. They can be applied to multislice, multiecho, water/fat, and gating schemes as well. Since motion problems are universal, it would appear that these modified sequences should come into common usage for MR imaging.

Humans↗